Peptide antagonists of the calcitonin cgrp family of peptide hormones and their use

Abstract

The embodiments provide a modified calcitonin gene-related peptide antagonist including an N-terminal fragment of modified calcitonin gene-related peptide or related protein family member where at least two residues of the N-terminal fragment are cysteine (Cys) and at least one amino acid comprises a non-threonine substitution of a threonine (Thr) residue; a central core where the central core comprises an a-helix; and a C-terminal fragment of modified calcitonin gene-related peptide or related protein family member comprising a C-terminal amide and where at least one amino acid of the C-terminal fragment is phenylalanine (Phe), proline (Pro), tyrosine (Tyr) or hydroxyproline (Hyp) or pharmaceutically acceptable salt thereof, as well as compositions, including pharmaceutical compositions, comprising a subject peptide. The embodiments further provide treatment methods, including methods of treating a migraine, the methods generally involving administering to an individual in need thereof an effective amount of a subject peptide or composition.

1 . A modified calcitonin gene-related peptide antagonist, or a pharmaceutically acceptable salt thereof, said antagonist having the structure of Formula I:
   X 1 —Y 1 —Z 1    (I)
   wherein:   X 1  is a modified N-terminal fragment of calcitonin gene-related peptide other comprising from five to seven amino acid residues, wherein two amino acid residues of the N-terminal fragment are cysteine (Cys), wherein the C-terminal amino residue of the fragment is cysteine (Cys), and wherein the residue immediately preceding the C-terminal Cys residue of the N-terminal fragment is a non-threonine substitution of threonine, wherein the two cysteine residues are separated by four, five or six amino acids and wherein the two cysteine residues can form a disulfide bond;   Y 1  is a central core comprising 15 to 22 amino acids wherein at least one amino acid of the central core is arginine (Arg) or lysine (Lys) and the central core comprises an a-helix; and   Z 1  is Val-Gly-Ser-Lys-Ala-Phe-NH 2 .   
     
     
         2 .- 45 . (canceled) 
     
     
         46 . The antagonist of  claim 1  wherein the central helical core comprises a fragment of human or salmon calcitonin. 
     
     
         47 . The antagonist of  claim 1 , wherein Y 1  comprises 18 to 20 amino acids. 
     
     
         48 . The antagonist of  claim 47 , wherein Y 1  is selected from the group consisting of -Val-Leu-Gly-Arg-Leu-Ser-Gln-Glu-Leu-His-Arg-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn- (SEQ ID NO: 34) or -Val-Leu-Gly-Lys-Leu-Ser-Gln-Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn- (SEQ ID NO: 35) or -Val-Leu-Gly-Lys-Leu-Ser-Gln-Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asp- (SEQ ID NO: 37) or -Val-Leu-Gly-Lys-Leu-Ser-Gln-Glu-Leu-His-Lys-Leu-Gln-Thr-Phe-Pro-Arg-Thr-Asn- (SEQ ID NO: 38) or -Val-Leu-Gly-Lys-Leu-Ser-Gln-Asp-Ile-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn- (SEQ ID NO: 39) or -Val-Leu-Gly-Lys-Leu-Ser-Gln-Glu-Leu-His-Lys-Met-Gln-Thr-Tyr-Pro-Arg-Thr-Asp (SEQ ID NO: 40) or -Leu-Leu-Gly-Lys-Leu-Ser-Gln-Glu-Leu-His-Arg-Leu-Gln-Thr-Tyr-Thr-Arg-Thr-Asp- (SEQ ID NO: 41) or -Val-Leu-Gly-Lys-Leu-Ser-Gln-Asp-Leu-His-Lys-Leu-Gln-Thr-Phe-Pro-Arg-Thr-Asp- (SEQ ID NO: 42) or -Met-Leu-Gly-Lys-Leu-Ser-Gln-Asp-Leu-His-Lys-Leu-Gln-Thr-Phe-Pro-Arg-Thr-Asp- (SEQ ID NO: 43) or -Val-Leu-Gly-Lys-Leu-Ser-Gln-Asp-Ile-His-Lys-Leu-Gln-Thr-His-Pro-Arg-Thr-Asp- (SEQ ID NO: 44). 
     
     
         49 . The antagonist of  claim 48  wherein Y 1  is -Val-Leu-Gly-Arg-Leu-Ser-Gln-Glu-Leu-His-Arg-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn- (SEQ ID NO: 34) or -Val-Leu-Gly-Lys-Leu-Ser-Gln-Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn- (SEQ ID NO: 35). 
     
     
         50 . The antagonist of  claim 49  wherein Y 1  has 95% sequence identity with V al-Leu-Gly-Arg-Leu-Ser-Gln-Glu-Leu-His-Arg-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-(SEQ ID NO 34) or -Val-Leu-Gly-Lys-Leu-Ser-Gln-Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn- (SEQ ID NO: 35). 
     
     
         51 . The antagonist of  claim 50  wherein Y 1  has 90% sequence identity with Val-Leu-Gly-Arg-Leu-Ser-Gln-Glu-Leu-His-Arg-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-(SEQ ID NO: 34). 
     
     
         52 . The antagonist of  claim 51  wherein Y 1  has 85% sequence identity with Val-Leu-Gly-Arg-Leu-Ser-Gln-Glu-Leu-His-Arg-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-(SEQ ID NO: 34). 
     
     
         53 . The antagonist of  claim 52  wherein Y 1  has 80% sequence identity with Val-Leu-Gly-Arg-Leu-Ser-Gln-Glu-Leu-His-Arg-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-(SEQ ID NO: 34). 
     
     
         54 . The antagonist of  claim 1 , wherein X 1  selected from the group consisting of NH 2 -Ala-Cys-Asp-Thr-Ala-Ala-Cys- (SEQ ID NO: 17), NH 2 -Ala-Cys-Asp-Thr-Ala-Ser-Cys- (SEQ ID NO: 18), NH 2 -Ala-Cys-Asp-Thr-Ala-Val-Cys- (SEQ ID NO: 19), NH 2 -Ala-Cys-Asn-Thr-Ala-Ala-Cys- (SEQ ID NO: 20), NH 2 -Ala-Cys-Val-Leu-Gly-Ala-Cys- (SEQ ID NO: 21), NH 2 -Ala-Cys-Arg-Phe-Gly-Ala-Cys- (SEQ ID NO: 22), NH 2 -Ala-Cys-Asp-Leu-Ser-Ala-Cys- (SEQ ID NO: 23), NH 2 -Ala-Cys-Asn-Leu-Ser-Ala-Cys- (SEQ ID NO: 24), NH 2 -Cys-Ser-Asn-Thr-Ala-Ala-Cys- (SEQ ID NO: 25), NH 2 -Ala-Cys-Asp-Thr-Ala-Leu-Cys- (SEQ ID NO: 26), NH 2 -Ala-Cys-Asp-Thr-Ala-Ile-Cys- (SEQ ID NO: 27), NH 2 -Ala-Cys-Asp-Thr-Ala-Leu-Cys- (SEQ ID NO: 28), NH 2 -Ala-Cys-Asp-Thr-Ala-Ile-Cys- (SEQ ID NO: 29), NH 2 -Ala-Cys-Asp-Leu-Ser-Val-Cys- (SEQ ID NO: 30), NH 2 -Ala-Cys-Asp-Leu-Ser-Val-Cys- (SEQ ID NO: 31), NH 2 -Ala-Cys-Asn-Leu-Ser-Val-Cys- (SEQ ID NO: 32), and NH 2 -Cys-Ser-Asn-Thr-Ala-Val-Cys- (SEQ ID NO: 33). 
     
     
         55 . A pharmaceutical composition comprising a pharmaceutically acceptable excipient and a modified calcitonin gene-related peptide antagonist of  claim 1 . 
     
     
         56 . A method of treating a condition associated with aberrant levels of CGRP comprising the administration of a modified calcitonin gene-related peptide antagonist of  claim 1 , to an individual, the method comprising administering to the individual an effective amount of a modified calcitonin gene-related peptide antagonist. 
     
     
         57 . The method of  claim 56 , wherein the condition is migraine. 
     
     
         58 . A method of treating a headache in an individual, the method comprising administering to the individual an effective amount pharmaceutical composition of  claim 55 . 
     
     
         59 . The method of  claim 58 , wherein the headache is migraine.